亚砷酸钠所致L-02人肝细胞损伤与p14ARF表达下调及MDM2、 p53表达增加有关  

Injury of L-02 hepatocytes induced by NaAsO is related to down-regulated expression of p14ARF and increased expression of MDM2 and p53

作　　者：王甜 赵哲仪 穆银贵 谢婷婷 罗昭逊 WANG Tian;ZHAO Zheyi;MU Yingui;XIE Tingting;LUO Zhaoxun(Department of Basic Clinical Laboratory Medicine,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学医学检验学院临床检验学教研室,贵州贵阳550004

出　　处：《细胞与分子免疫学杂志》2020年第6期507-512,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81560514);贵州省卫生计生委科学技术基金(gzwjkj2014-1-026);大学生创新创业训练计划(201510660004)。

摘　　要：目的研究亚砷酸钠(NaAsO2)致L-02人肝细胞损伤过程中p14可变读框(p14ARF)、鼠双微基因2(MDM2)和p53的表达变化。方法用(0、5、10、15、20、25)μmol/L NaAsO2分别处理L-02细胞48 h,相差显微镜观察细胞形态变化,CCK-8法检测细胞增殖,异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞凋亡,实时荧光定量PCR和Western blot法分别检测p14ARF、MDM2和p53 mRNA及蛋白表达。结果与对照组相比,随着NaAsO2浓度增加,染砷组L-02肝细胞增殖活性逐渐降低,细胞凋亡率逐渐升高,p14ARF mRNA及蛋白水平逐渐降低,MDM2 mRNA及蛋白水平逐渐升高,p53 mRNA水平变化不明显,p53蛋白水平逐渐升高。结论NaAsO2所致L-02肝细胞损伤可能与p14ARF表达下调及p53、MDM2表达的上调有关。Objective To investigate the effects of sodium arsenite(NaAsO2)on p14 alternative reading frame(p14ARF),murine double minute 2(MDM2)and p53 expressions in L-02 hepatocytes.Methods L-02 hepatocytes were cultured in medium containing(0,5,10,15,20,25)μmol/L NaAsO2 for 48 hours.The cell morphology was evaluated by phase-contrast microscopy and cell proliferation was detected by CCK-8 assay.The apoptosis of hepatocytes was detected by annexinⅤ/propidium iodide(annexinⅤ-FITC/PI)double staining followed by flow cytometry.The mRNA and protein expressions of p14ARF,MDM2 and p53 were detected by real-time fluorescence quantitative PCR and Western blotting.Results Compared with the control group,the proliferation activity of L-02 hepatocytes decreased and the apoptotic rate of L-02 hepatocytes increased significantly.With the increase of NaAsO2 concentration,p14ARF mRNA and protein levels decreased gradually,while MDM2 mRNA and protein levels increased gradually.There was no significant difference in the expression of p53 mRNA,but the relative expression level of p53 protein increased gradually.Conclusion The injury of L-02 hepatocytes induced by NaAsO2 may be related to the down-regulation of p14ARF expression and the up-regulation of p53 and MDM2 expression.

关 键 词：亚砷酸钠(NaAsO2) L-02肝细胞 p14可变读框(p14ARF) 鼠双微基因2(MDM2) P53 

分 类 号：R994.6[医药卫生—毒理学] R122[医药卫生—药学]